Cloning pigs using donor nuclei from non-quiescent differentiated cells
AssigneeUniversity of Massachusetts
InventorsSteven L. Stice, James M. Robl, Jose Cibelli, Paul Golueke
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Abstract
An improved method of nuclear transfer involving the transplantation of donor differentiated pig cell nuclei into enucleated pig oocytes is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring. Production of genetically engineered or transgenic pig embryos, fetuses and offspring is facilitated by the present method since the differentiated cell source of the donor nuclei can be genetically modified and clonally propagated.
Claims26
A method of cloning a pig, comprising:
- (i) inserting a non-quiescent differentiated pig cell or non-quiescent differentiated pig cell nucleus into an enucleated pig oocyte, under conditions suitable for the formation of a nuclear transfer (NT) unit;
- (ii) activating the resultant NT unit;
- and
- (iii) transferring said activated NT unit to a host pig such that the NT unit develops into a fetus.
A method of cloning a pig, comprising:
- (i) inserting a non-quiescent differentiated pig CICM cell or non-quiescent differentiated pig CICM cell nucleus into an enucleated pig oocyte, under conditions suitable for the formation of a nuclear transfer (NT) unit;
- (ii) activating the resultant NT unit;
- and
- (iii) transferring said activated NT unit to a host pig such that the NT unit develops into a fetus.
A method of producing a non-human mammalian embryo by nuclear transfer comprising transplantation of a non-human mammalian cell or a nucleus of a non-human mammalian cell into an enucleated oocyte of the same species as the donor cell or donor cell nucleus, activation of the recipient oocyte containing the donor cell or donor cell nucleus, and incubation of the activated oocyte to produce an embryo, wherein the donor cell is a non-quiescent mammalian differentiated cell or wherein the donor nucleus is from a non-quiescent mammalian differentiated cell.
Dependencies are inferred from the claim text.